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S A Ward

Publications and source records attributed to S A Ward.

At least 91 records · Page 5Linked to original sources
PubMed1996

The role of drug accumulation in 4-aminoquinoline antimalarial potency. The influence of structural substitution and physicochemical properties.

We have investigated a series of novel 4-aminoquinoline analogues related to amodiaquine, that possess side chain modifications designed to influence both drug pKa and lipophilicity. These compounds have been used to determine the influence of physicochemical properties on antimalarial activity against, and accumulation by, both chloroquine-susceptible and chloroquine-resistant isolates of Plasmodium falciparum. The compounds tested exhibited a 500-fold range of absolute antimalarial potency. Absolute drug potency and drug accumulation were found to be significantly correlated in each of the four isolates of Plasmodium falciparum studied. The level of accumulation was unrelated to lipophilicity and was significantly greater than the predicted levels of accumulation based on drug pKa, compartmental pH, and Henderson-Hasselbach considerations. Further analysis of the relationship between 4-aminoquinoline accumulation and activity implicated the involvement of additional forces in the accumulation process.

Aminoquinolines↗
PubMed1996

Amodiaquine accumulation in Plasmodium falciparum as a possible explanation for its superior antimalarial activity over chloroquine.

Amodiaquine is a 4-aminoquinoline antimalarial whose structure is similar to chloroquine. In contrast to the wealth of information available about chloroquine accumulation and its relationship to activity, little is known about the uptake characteristics of amodiaquine, a drug that is inherently more active against malaria parasites. In this study we have investigated the accumulation of amodiaquine in Plasmodium falciparum in vitro, in order to gain an insight into the mechanisms responsible for its superior activity over chloroquine. The driving force for parasite accumulation of the 4-aminoquinolines is proposed to be a transmembrane proton gradient maintained by a vacuolar ATPase. In the present study, amodiaquine accumulation was greatly reduced, at steady state, in the absence of glucose and at 0 degrees C indicating a clear energy dependence of uptake. Amodiaquine accumulation in Plasmodium falciparum was shown to be 2- to 3-fold greater than chloroquine accumulation. This observation probably accounts for amodiaquine's greater inherent activity but is surprising given that amodiaquine is a weaker base than chloroquine. With this in mind we present evidence for an intraparasitic binding component in the accumulation of the 4-aminoquinolines. Differences in binding affinity of this 'receptor' for amodiaquine and chloroquine may partially explain the greater accumulation and in vitro potency of amodiaquine compared to chloroquine.

Adenosine Triphosphate↗
PubMed1996

Manipulation of the N-alkyl substituent in amodiaquine to overcome the verapamil-sensitive chloroquine resistance component.

Aminoquinoline resistance correlates with lipid solubility at pH 7.2. Consequently, the in vivo dealkylation of amodiaquine, to the less lipid-soluble desethylamodiaquine, is a likely contributor to therapeutic failure in vivo. Therefore, 4-aminoquinoline drugs with lipid solubilities similar to that of amodiaquine, but which are not subject to side chain modification in vivo, should be superior antimalarial agents. In this study, we have identified amopyroquine and N-tertbutylamodiaquine as two such compounds. The values for the logarithms of the partition coefficients for amopyroquine and N-tertbutylamodiaquine are between those for amodiaquine and its dealkylated metabolite, desethylamodiaquine. Both amopyroquine and N-tertbutylamodiaquine possess levels of antimalarial activity greater than that of desethylamodiaquine and significantly reduced cross-resistance patterns; i.e., the former two compounds are not subject to the verapamil-sensitive resistance mechanism. Simple in vitro markers of direct toxicity and potential reactive metabolite formation suggest that these two compounds are no more toxic than amodiaquine and desethylamodiaquine.

Alkylation↗
PubMed1996

Effects of prior exercise on pulmonary gas-exchange kinetics during high-intensity exercise in humans.

The effects of prior exercise on O2 uptake (VO2) kinetics during supra-lactate threshold (LT) cycling were assessed in 11 subjects. Protocols consisted of two consecutive 6-min work bouts separated by 0 W (6 min) with 1) both bouts sub-LT, 2) both bouts supra-LT, 3) bout 1 sub-LT and bout 2 supra-LT, and 4) bout 1 supra-LT and bout 2 sub-LT. Sub-LT VO2 kinetics were similar whether the prior bout was supra- or sub-LT. The VO2 kinetics for supra-LT work preceded by a sub-LT "warm-up" were similar to those for supra-LT work that was not preceded by exercise (O-W warm-up): the "partial": O2 deficit averaged 2.64 vs. 2.57 liters, and the "effective" VO2 time constant averaged 56 vs. 65 s. Exercise responses (i.e., the change between O W and minute 6 of exercise) were unaffected for lactate concentration (4.58 vs. 4.50 meq/l), pH (-0.08 vs. -0.10), and CO2 output (VCO2; 2.65 vs. 2.49 l/min). However, when the supra-LT work was preceded by a supra-LT warm-up, VO2 kinetics were appreciably faster (O2 deficit = 1.82 liters, VO2 time constant = 37 s) relative to 0-W warm-up; the lactate (0.69 meq/l), pH (-0.01), and VCO2 (2.08 l/min) responses were smaller; and the effective VCO2 time constant was longer (58 vs. 43 s). The mechanism(s) that underlie this speeding of the VO2 kinetics cannot be firmly established, but we suggest that an improved muscle perfusion during the exercise may be involved consequent to the residual metabolic acidemia from the high-intensity warm-up.

Acidosis↗
PubMed1996

4-Aminoquinoline resistance of Plasmodium falciparum: insights from the study of amodiaquine uptake.

The relationship between antimalarial activity and drug accumulation of chloroquine and amodiaquine was evaluated with four chloroquine-resistant and two chloroquine-susceptible isolates of Plasmodium falciparum. Susceptibility of the strains to amodiaquine was correlated with susceptibility to chloroquine (r2 = 0.96). Similarly, accumulation of amodiaquine was correlated with accumulation of chloroquine (r2 = 0.94). Accumulation of both chloroquine and amodiaquine was significantly reduced in chloroquine-resistant isolates (p < 0.005). For the panel of isolates, the accumulation ratio of both drugs was inversely proportional to drug susceptibility (r2 = 0.963 and 0.994 for amodiaquine and chloroquine, respectively). Time course studies highlighted a reduced initial rate of amodiaquine accumulation in chloroquine-resistant isolates compared with chloroquine-susceptible isolates, with no evidence of an enhanced drug efflux rate. Daunomycin, a modulator of parasite chloroquine transport, significantly increased steady state accumulation of both drugs in chloroquine-resistant isolates and, to a lesser extent, in chloroquine-susceptible isolates. Furthermore, daunomycin increased the initial rate of accumulation of amodiaquine in both chloroquine-resistant and chloroquine-susceptible isolates. Resistance to 4-aminoquinoline drugs is associated with reduced drug permeability rather than enhanced cellular exit of preaccumulated drug, and daunomycin seems to increase the permeability of parasites to aminoquinolines. A new model of 4-aminoquinoline resistance is proposed to take account of these and earlier observations.

Aminoquinolines↗
PubMed1996

Physicochemical properties correlated with drug resistance and the reversal of drug resistance in Plasmodium falciparum.

At high molar excess, verapamil can selectively increase the accumulation and cytotoxicity of structurally dissimilar natural product drugs in many multidrug-resistant tumor cell lines. Such concentrations of verapamil are also capable of increasing the accumulation and activity of chloroquine in chloroquine-resistant strains of the human malaria parasite Plasmodium falciparum. Despite such similarities, it is not clear why chloroquine-resistant P. falciparum is often susceptible to closely related compounds such as amodiaquine, whereas cancer cells are cross-resistant to many structurally unrelated drugs. For 13 aminoquinoline and aminoacridine compounds, relative drug resistance was negatively correlated with lipid solubility at physiological pH (r2 = 0.90, p < 0.0001). The ability of verapamil (5 microM) to reverse drug resistance was also negatively correlated with lipid solubility (r2 = 0.88, p < 0.0001). Furthermore, molar refractivity was weakly correlated with relative drug resistance (r2 = 0.46, p < 0.05) and reversal of drug resistance (r2 = 0.52, p < 0.005). Verapamil increases chloroquine accumulation by resistant parasites, a mechanism suggested to account for its selective chemosensitization effect. We show that the initial rate of chloroquine accumulation by resistant parasites is increased by verapamil. This effect of verapamil is abolished when deoxy-glucose is substituted for glucose. Therefore, verapamil produces an energy-dependent increase in the permeability of resistant parasites to chloroquine. For a panel of four chloroquine-resistant and two chloroquine-susceptible isolates, the effect of verapamil on the accumulation of chloroquine and monodesethyl amodiaquine was found to be correlated (r2 = 0.96, p < 0.001). Verapamil chemosensitization was also correlated for the two drugs (r2 = 0.92, p < 0.005), suggesting a common mechanism. In summary, the degree of drug resistance and the extent of verapamil chemosensitization for a particular drug seem to be dependent on general physical features such as lipid solubility and molar refractivity rather than on closely defined structural parameters. These studies provide insight into this important resistance mechanism of malaria parasites and may provide direction for the development of new drugs that are effective against resistant parasites.

Animals↗
PubMed1995

The toxicity of artemisinin and related compounds on neuronal and glial cells in culture.

The antimalarial drug artemisinin and a number of its derivatives were tested for their effects on proliferation of undifferentiated neuroblastoma Nb2a cells and glioma C6 cells in culture as well as their ability to inhibit neurite outgrowth from Nb2a cells differentiated by removal of serum and addition of dibutyryl cyclic AMP. In the Nb2a and C6 cell cultures, all drugs except desoxyartemisinin significantly inhibited cell proliferation in a dose-related manner with the lowest effective concentration being that of artemisinin at 0.1 microM. Artemether, arteether, artemisinin and dihydroartemisinin also produced a dose-related decrease in the number of neurites/extensions formed by differentiating Nb2a cells, with an effect of dihydroartemisinin at a concentration as low as 1 nM. Desoxyartemisinin had no effect on extension/neurite formation. We propose a potential mechanism for neurotoxicity of artemisinin and its derivatives that involves the endoperoxide bridge which is also known to be necessary for their antimalarial action.

Animals↗
PubMed1995

Obligatory anaerobiosis resulting from oxygen uptake-to-blood flow ratio dispersion in skeletal muscle: a model.

We constructed a computer model whereby the dispersion of the oxygen uptake-to-blood flow ratio (VO2/Q) in working muscle could be altered around its mean level during simulated exercise. The model incorporated standard values for the cardiac output and whole-body O2 uptake responses to incremental exercise in humans. Dispersion of the VO2/Q ratio was induced by distributing Q in a Gaussian fashion around its appropriate mean value with a standard deviation (sigma). By increasing sigma, more regions of the muscle became "anaerobic" as the corresponding local muscle venous blood O2 content fell to zero; these regions thus required proportionally greater rates of lactate production to sustain the required energy transfer. The model yielded a relatively high mixed-muscle venous partial pressure of O2, suggesting that the latter may not be a sufficient indicator of whether or not there may be diffusional limitation to muscle O2 transfer during exercise or whether or not there may be anaerobic production of lactate in certain regions of exercising muscle.

Anaerobiosis↗
PubMed1995

Current views on the mechanisms of resistance to quinoline-containing drugs in Plasmodium falciparum.

The issue of chloroquine resistance in Plasmodium falciparum and cross-resistance patterns with other related chemotherapeutic agents has been the subject of intense interest for many years. Despite this level of investigation, the picture remains very unclear. Although it is accepted that chloroquine resistance is, at least in part, a function of reduced drug accumulation, the question of reduced drug uptake versus enhanced efflux is yet to be resolved at both the molecular and biochemical levels. Further, the absolute cross-resistance patterns of chloroquine-resistant isolates to closely related analogues is a matter for debate, although there appears to be a reciprocal arrangement between resistance to chloroquine and resistance to mefloquine, halofantrine and possibly quinine. Evidence is presented for the coexistence of two or more chloroquine-resistance mechanisms in isolates of P. falciparum, only one of which is verapamil sensitive. In addition, an analysis of cross-resistance patterns, as measured by the inoculum effect, is presented.

Animals↗
PubMed1995

The role of S-mephenytoin hydroxylase (CYP2C19) in the metabolism of the antimalarial biguanides.

The effects of the CYP2C19 substrates, mephenytoin, methsuximide and mephobarbitone on the metabolism of proguanil and chlorproguanil by human liver microsomes were studied. All of the CYP2C19 substrates significantly inhibited (P < 0.05) the formation of both cycloguanil and chlorcycloguanil from their parent compounds. In the presence of mephenytoin (50 and 100 microM) the extent of proguanil cyclisation was decreased by 66% and 67% whilst the cyclisation of chlorproguanil was decreased by 51% and 70%, respectively. Methsuximide (50 and 100 microM) inhibited cycloguanil formation by 68% and 77% and chlorcycloguanil formation by 43% and 58%, respectively. In the presence of mephobarbitone (50 and 100 microM) the cyclisation of proguanil and chlorproguanil to their active metabolites was reduced by 24% and 42% and 48% and 63%, respectively. In addition, proguanil and chlorproguanil were shown to be mutual competitive inhibitors of metabolism to their triazine metabolites. In the presence of proguanil (50 and 100 microM) the Km value for chlorcycloguanil production was increased by 118% and 200%, respectively, with little change in Vmax. Similarly, chorproguanil (50 microM) increased the Km for the in vitro cyclisation of proguanil by 50% with no alteration in Vmax. These data suggest that both chlorproguanil and proguanil are metabolised in vitro by mephenytoin hydroxylase, CYP2C19.

Anticonvulsants↗
PubMed1995

The effect of chemical substitution on the metabolic activation, metabolic detoxication, and pharmacological activity of amodiaquine in the mouse.

The adverse reactions associated with the antimalarial amodiaquine (AQ), agranulocytosis and hepatotoxicity, have been attributed to the bioactivation of the drug to a quinone imine metabolite. Therefore the effect of chemical modification on the metabolism of AQ was studied, with particular reference to the prevention of bioactivation and the introduction of glucuronidation. Glutathione conjugates of AQ and desethylAQ were eliminated in bile after intraportal administration of [3H]AQ (54 mumol/kg, 20 microCi/kg) to anesthetized male CD1 mice. Thioether conjugates excreted into bile over 3 h accounted for 28% of the administered dose. Fluorine substitution at the C-4 position of AQ blocked bioactivation, as measured by formation of thioether conjugates, and resulted in a 5-fold decrease in biliary excretion of radiolabeled dose: ca 6% versus ca 29%. Additional substitution of a primary alcohol function into one of the ethyl moieties introduced glucuronidation as a pathway of elimination, with 10% of the dose being excreted in bile as an O-glucuronide of the parent compound over a 3-h period; excretion of total radioactivity in bile increased 2.5-fold. These substitutions resulted in a 2-fold greater excretion of radiolabel into urine: 41% and 39% for DFAQ and HDFAQ, respectively, versus 23% for AQ. Novel carboxylic acid and N-oxide metabolites of the fluorinated analogues were identified. AQ and the two fluorinated analogues had similar activity against Plasmodium berghei in mice. These results demonstrate that the metabolism of AQ can be diverted from extensive bioactivation to direct detoxication by simple chemical substitutions that do not impair pharmacological activity.

Amodiaquine↗
PubMed1994

Ferriprotoporphyrin catalysed decomposition of artemether: analytical and pharmacological implications.

The ability of the products of erythrocytic haemolysis and ferriprotoporphyrin (FP-IX)-containing substances of facilitate the decomposition of the antimalarial drug artemether has been evaluated in vitro. The products of haemolysis accelerate the degradation of artemether to unidentified compounds which are undetectable by currently available HPLC methods. This decomposition is temperature dependent and occurs after relatively brief artemether (ARM)-catalyst contact. Using radiolabelled [16-14C]ARM, we have demonstrated that the extractability of total radioactivity into the organic phase diminishes with increasing FP-IX concentration with an appreciable reduction in the organic extractability as ARM in the presence of FP-IX and associated increase in water solubility of radioactivity when the concentration of FP-IX increases from 0 to 7.7 mM. This effect appears to be temperature dependent for incubations with haematin. Characterisation of the organically extractable radioactivity from incubations of [16-14C]ARM with FP-IX has shown a loss of ARM and the formation of two radioactive products which occurs in proportion to the concentration of FP-IX. We believe these findings are of particular relevance to the determination of plasma concentrations of ARM and dihydroartemisinin (DHA) in malaria patients where extensive haemolysis and the presence of breakdown products of haemoglobin may contribute to a reduction in the circulating concentration of these substances. In these circumstances, measurement of ARM and DHA by conventional methods may be meaningless.

Antimalarials↗
PubMed1994

The effect of fluorine substitution on the metabolism and antimalarial activity of amodiaquine.

Amodiaquine (AQ) (2) is a 4-aminoquinoline antimalarial which causes adverse side effects such as agranulocytosis and liver damage. The observed drug toxicity is believed to be related to the formation of an electrophilic metabolite, amodiaquine quinone imine (AQQI), which can bind to cellular macro-molecules and initiate hypersensitivity reactions. 5'-Fluoroamodiaquine (5'-FAQ, 3), 5',6'-difluoroamodiaquine (5',6'-DIFAQ,4), 2',6'-difluoroamodiaquine (2',6'-DIFAQ,5), 2',5',6'-trifluoroamodiaquine (2',5',6'-TRIFAQ, 6) and 4'-dehydroxy-4'-fluoroamodiaquine (4'-deOH-4'-FAQ,7) have been synthesized to assess the effect of fluorine substitution on the oxidation potential, metabolism, and in vitro antimalarial activity of amodiaquine. The oxidation potentials were measured by cyclic voltammetry, and it was observed that substitution at the 2',6'- and the 4'-positions (2',6'-DIFAQ and 4'-deOH-4'-FAQ) produced analogues with significantly higher oxidation potentials than the parent drug. Fluorine substitution at the 2',6'-positions and the 4'-position also produced analogues that were more resistant to bioactivation. Thus 2',6'-DIFAQ and 4'-deOH-4'-FAQ produced thioether conjugates corresponding to 2.17% (SD: +/- 0.27%) and 0% of the dose compared with 11.87% (SD: +/- 1.31%) of the dose for amodiaquine. In general the fluorinated analogues had similar in vitro antimalarial activity to amodiaquine against the chloroquine resistant K1 strain of Plasmodium falciparum and the chloroquine sensitive T9-96 strain of P. falciparum with the notable exception of 2',5',6'-TRIFAQ (6). The data presented indicate that fluorine substitution at the 2',6'-positions and replacement of the 4'-hydroxyl of amodiaquine with fluorine produces analogues (5 and 7) that maintain antimalarial efficacy in vitro and are more resistant to oxidation and hence less likely to form toxic quinone imine metabolites in vivo.

Amodiaquine↗
PubMed1994

Relationship of global chloroquine transport and reversal of resistance in Plasmodium falciparum.

Control of falciparum malaria has become almost impossible in many areas due to the development of resistance to chloroquine and other antimalarial drugs. Verapamil and a number of unrelated compounds which chemosensitise multi-drug resistant cancer cells also enhance chloroquine susceptibility in Plasmodium falciparum. Chloroquine is accumulated to lower levels in resistant plasmodia, hence the reversal of chloroquine resistance has been attributed to the ability of chemosensitising agents to increase the amount of chloroquine accumulated by the resistant parasite. We have conducted a detailed examination of the effect of verapamil on chloroquine sensitivity and its relationship to chloroquine accumulation. The ability of verapamil to increase steady-state chloroquine accumulation was found to be totally insufficient to explain the increase in chloroquine sensitivity caused by the drug. In contrast, when chloroquine accumulation was increased by raising the pH gradient, the corresponding shifts in sensitivity to chloroquine could be accurately predicted. These results were confirmed with other classes of chemosensitisers and we conclude that an alternative mechanistic explanation is required to completely explain the reversal of chloroquine resistance in P. falciparum.

Animals↗
PubMed1994

Pharmacokinetics of artemether after oral administration to healthy Thai males and patients with acute, uncomplicated falciparum malaria.

1. The pharmacokinetics of artemether were investigated (a) in six healthy male Thai volunteers after single 200 mg oral doses and (b) in eight male Thai patients with acute uncomplicated falciparum malaria after an initial 200 mg oral dose followed by 100 mg at 12 h then 100 mg daily for 4 days. 2. In the healthy subjects, median (range) maximum plasma concentrations of artemether of 118 (112-127) ng ml-1 were reached at 3 (1-10) h. Thereafter, drug concentrations declined monoexponentially with a median (range) t1/2.z of 3.1 (1.0-9.6) h. The median (range) AUC and MRT values were 1.10 (0.33-4.44) micrograms ml-1 h and 8.3 (3.5-20.8) h. The median Cmax value of dihydroartemisinin, an active metabolite, was 379 (162-702) mg ml-1 at 6 (2-12) h. Its median AUC value was 6.6 (0.83-38.7) micrograms ml-1 h; the apparent t1/2.z was 10.6 (4.7-19.2) h and the median MRT value was 16.0 (5.0-41.0) h. 3. In the patients, a higher Cmax value of parent drug than those observed in healthy subjects (median and range of 231 (116-411) ng ml-1), was reached at 3 (1-3) h after the first dose. Steady state was reached after the third dose (24 h) and concentrations fluctuated over the range of 36-60 ng ml-1. The respective median (range) values of AUC and t1/2.z were 5.8 (3.76-12.9) micrograms ml-1 h and 4.2 (2.5-5.3) h.(ABSTRACT TRUNCATED AT 250 WORDS)

Administration, Oral↗
PubMed1994

Peripheral and central chemoreceptor control of ventilation during exercise in humans.

The stability of arterial blood gas tensions and pH during steady-state moderate exercise has suggested an important humoral element of ventilatory control in humans. However, the involvement of central and peripheral chemoreflexes in this humoral control remains controversial. This reflects, in large part, technical and interpretational limitations inherent in currently used estimators of chemoreflex "sensitivity." Evidence suggests that the central chemoreceptors (a) contribute little during moderate exercise, given the relative stability of cerebrospinal pH, (b) constrain the hyperpnea of high-intensity exercise, consequent to the respiratory compensation for the metabolic acidemia, and (c) may play a role in the respiratory compensation during chronic metabolic acidemia. In contrast, the peripheral chemoreceptors appear to (a) exert considerable influence on ventilatory kinetics in moderate exercise, but are less important in the steady state, and (b) induce much of the respiratory compensation of high-intensity exercise.

Acidosis↗
PubMed1994

Effects of infused epinephrine on slow phase of O2 uptake kinetics during heavy exercise in humans.

We tested the hypothesis that infused epinephrine (Epi) would augment the slow phase of oxygen uptake (VO2) during heavy exercise. Six normal healthy males initially performed a ramp test on a cycle ergometer to estimate the lactate threshold (LT) and determine peak VO2. Each subject then performed two 20-min constant-load tests at a power output calculated to elicit a VO2 equal to estimated LT + 0.2(peak VO2--estimated LT) under control conditions throughout and with an intravenous infusion of Epi from minutes 10 to 20 at a rate of 100 ng.kg-1.min-1. Pulmonary gas exchange variables were determined breath by breath. Arterialized venous blood was repeatedly sampled from the dorsum of the heated hand. Epi infusion elevated (P < 0.05) plasma Epi concentration (i.e., from 420 +/- 130 pg/ml at minute 10 to 2,190 +/- 410 pg/ml at minute 20) but had no effect on plasma norepinephrine or K+ concentrations. Concentrations of blood lactate and pyruvate were increased, pH was decreased, and base excess became more negative by infusion of Epi (P < 0.05). Epi infusion increased (P < 0.05) CO2 production and the respiratory exchange ratio but had no effect on ventilation or VO2. VO2 increased (P < 0.05) to the same extent in both control (3.14 +/- 0.12 l/min at minute 10, 3.28 +/- 0.12 l/min at minute 20) and Epi infusion (3.10 +/- 0.11 l/min at minute 10, 3.25 +/- 0.11 l/min at minute 20) trials. We therefore concluded that neither Epi nor its associated humoral consequences contribute significantly to the slow phase of VO2 kinetics during heavy exercise.

Adult↗